Method for detecting target sequences

A method for detecting one or more target sequences in a sample comprising RNA by amplification, the method comprising:

• a) providing an aqueous composition comprising:
• i) one or more forward oligonucleotide primers having a 5′ region which is a tag sequence and a 3′ region specific for a target sequence;
• ii) a reverse oligonucleotide primer, such that a forward and the reverse primer operably form a primer pair;
• iii) one or more probes having a reporter label and a quencher label, each probe comprising a sequence having at least 50% identity to the tag sequence of a forward oligonucleotide primer;
• and
• iv) optionally one or more enhancer oligonucleotide primers comprising a sequence having at least 50% identity to the sequence of a probe;
• b) combining the primers, the probes, the sample, a polymerase, and a reverse transcriptase in a reaction mixture with a buffer;
• and
• c) performing a one-step polymerase chain reaction (PCR) on the reaction mixture to generate tagged nucleic acids, whereby the probe(s) can bind the tagged nucleic acids.